The Hevylite IgG Kappa kit is a quantitative in vitro assay intended for the measurement of IgG Kappa (IgG heavy chain and Kappa light chain intact immunoglobulin) in serum using the Optilite analyser. Measurement of Hevylite IgG Kappa is used alongside Hevylite IgG Lambda to calculate the IgG Lambda ratio. The Hevylite IgG Kappa / IgG Lambda ratio can be used when monitoring previously diagnosed IgG multiple myeloma patients and is used in conjunction with other laboratory tests and clinical evaluations. The assignment of complete response is reliant upon other tests including immunofixation, bone marrow and urine assessments.

The Hevylite IgG Lambda kit is a quantitative in vitro assay intended for the measurement of IgG heavy chain and Lambda light chain intact immunoglobulin) in serum using Site Optilite analyser. Measurement of Hevylite IgG Lambda is used alongside Hevylite IgG Kappa to calculate the IgG Kappa / IgG Lambda ratio. The Hevylite IgG Kappa / IgG Lambda ratio can be used when monitoring previously diagnosed IgG multiple myeloma patients and is used in conjunction with other laboratory tests and clinical evaluations. The assignment of complete response is reliant upon other tests including immunofixation, bone marrow and urine assessments.

Device Description

The Hevylite Human IgG Kappa and IgG Lambda Kits contain vials of ready-to-use polyclonal monospecific sheep anti-lgG antisera against combined γ heavy and κ light chain or combined y heavy and À light chain, calibrators (six levels), controls (low and high) and reaction buffer in liquid form. The reagents contain 0.099% sodium azide as preservative.

AI/ML Overview

The provided text describes the performance characteristics of the Optilite Hevylite IgG Kappa Kit and Optilite Hevylite IgG Lambda Kit, which are in vitro diagnostics. It does not describe an AI medical device, but rather a laboratory assay. Therefore, many of the requested points, such as "multi reader multi case (MRMC) comparative effectiveness study," "standalone (i.e. algorithm only without human-in-the-loop performance)," and information on experts for ground truth and adjudication methods, are not applicable to this type of device.

However, I can extract and present the relevant information regarding the performance study of this In Vitro Diagnostic (IVD) assay.

Device Information:

Trade/Device Name: Optilite Hevylite IgG Kappa Kit, Optilite Hevylite IgG Lambda Kit
Regulation Number: 21 CFR 866.5510
Regulation Name: Immunoglobulins A, G, M, D, and E immunological test system
Regulatory Class: Class II
Product Code: PCN, PCO
Intended Use: Quantitative in vitro assay for the measurement of IgG Kappa (IgG heavy chain and kappa light chain intact immunoglobulin) and IgG Lambda (IgG heavy chain and lambda light chain intact immunoglobulin) in serum. Used alongside each other to calculate the IgG Kappa/IgG Lambda ratio, which can be used when monitoring previously diagnosed IgG multiple myeloma patients in conjunction with other laboratory tests and clinical evaluations.

Study Proving Device Meets Acceptance Criteria:

The study evaluated the analytical and clinical performance of the device by comparing it to a legally marketed predicate device (Hevylite Human IgG Kappa Kit and Hevylite Human IgG Lambda Kit for use on the Siemens BN™II, K132555).

1. Table of Acceptance Criteria and Reported Device Performance

The document does not explicitly list pre-defined "acceptance criteria" in a table format with pass/fail values. Instead, it presents the results of several performance studies which, taken together, demonstrate the device's suitability for its intended use and its substantial equivalence to the predicate device. The performance is demonstrated through precision, linearity, stability, detection limits, analytical specificity (interference and cross-reactivity), and method comparison studies. Clinical validity is supported by a monitoring response category comparison and data modeling.

Here's a summary of the performance metrics reported:

Performance Metric	Acceptance Criteria (Implied)	Reported Device Performance
Precision (Total %CV)	Acceptable coefficients of variation for different concentration levels.	IgGL Precision Studies:Sample 1 (4.260): 7.6%Sample 2 (3.148): 5.4%Sample 3 (5.601): 7.7%Sample 4 (7.621): 6.8%Sample 5 (13.574): 7.2%Sample 6 (16.824): 6.7%IgGK Precision Studies:Sample 1 (2.894): 9.0%Sample 2 (1.313): 6.2%Sample 3 (2.379): 4.2%Sample 4 (4.262): 7.5%Sample 5 (6.657): 5.3%Sample 6 (13.941): 4.0%
Linearity/Reportable Range	Linear over the standard measuring ranges with acceptable deviation from linearity.	IgGK: Linear over 1.930 - 33.427 g/L (at 1+19 dilution) with deviation from linearity ≤ 10%.IgGL: Linear over 1.380 - 19.300 g/L (at 1+19 dilution) with deviation from linearity ≤ 10%. (Standard measuring ranges: IgGK: 2.3 – 30.0 g/L, IgGL: 1.5 – 17.5 g/L).
Open Vial Stability	Maintain performance for specified duration.	Supports a claim of 3 months at 2-8°C.
On-Board Stability	Maintain performance for specified duration when on the instrument.	Supports a claim of 28 days (provided power is left switched on).
Detection Limits (LoD, LoQ)	Low enough for clinical utility.	IgG Kappa: LoB=0.000 g/L, LoD=0.009 g/L, LoQ=0.115 g/LIgG Lambda: LoB=0.000 g/L, LoD=0.005 g/L, LoQ=0.075 g/L
Analytical Specificity (Interference)	Interference should be within an acceptable range (e.g., ±10% deviation).	No significant assay interference effects observed with bilirubin (200mg/L), hemoglobin (5g/L), triglyceride (1000mg/dL), intralipid (125mg/dL), or 16 commonly used drugs at specified concentrations.
Antigen Excess Detection	Ability to detect antigen excess to prevent false low results.	No antigen excess observed up to 100.5 g/L for IgG Kappa and 102.5 g/L for IgG Lambda (concentrations above respective standard measuring ranges).
Method Comparison (Correlation with Predicate)	Strong correlation coefficient (r) and acceptable Passing Bablok regression equation parameters.	IgG Kappa: 284 serum samples (range 0.2 - 47.69 g/L). y = 1.10x - 0.68 g/L; r = 0.957 (y = Optilite, x = predicate analyser).IgG Lambda: 172 serum samples (range 0.09 - 40.65 g/L). y = 0.95x + 0.00 g/L; r = 0.823 (y = Optilite, x = predicate analyser).IgG Kappa/Lambda Ratio: 143 serum samples (range 0.01 - 277.50 g/L). y = 1.12x - 0.17; r = 0.901 (y = Optilite, x = predicate analyser).
Clinical Monitoring Response Category Agreement	High kappa/weighted kappa statistics indicating good agreement between device and predicate for classifying patient responses (CR, VGPR, PR, SD, PD).	Optilite Monitoring Study (69 samples): Kappa (95% CIs) = 0.79 (0.62 – 0.96); Weighted Kappa (95% CIs) = 0.87 (0.74 – 0.98). (For 43 response classifications out of 69 monitoring samples).Data Modelling (Transformed Data Response - H/L, 437 evaluations):* Kappa (95% CIs) = 0.75 (0.70 – 0.80); Weighted Kappa (95% CIs) = 0.86 (0.82 – 0.89).Data Modelling (Transformed Data Response - L/H, 437 evaluations):** Kappa (95% CIs) = 0.81 (0.76 - 0.85); Weighted Kappa (95% CIs) = 0.90 (0.86 - 0.93).
Reference Intervals	Verification of transferred reference intervals.	Verified by testing 50 adult donor samples. IgG kappa (g/L): 4.03 – 9.78 g/L; IgG lambda (g/L): 1.97 - 5.71 g/L; IgG kappa/ IgG lambda ratio: 0.98 - 2.75.

2. Sample Sizes Used for the Test Set and Data Provenance

Precision/Reproducibility: Six serum samples (contents not detailed, likely pooled or commercial controls/spiked samples) tested over 21 days with two runs per day on three different reagent lots on three analyzers.
Linearity: Serum samples (quantity not explicitly stated, but "serially diluted serum samples") to cover the standard measuring ranges.
Open-vial stability: Three lots of kits.
On-board stability: Three lots of kits.
Detection limit: Analyte depleted samples (60 determinations for LoB), 5 independent samples diluted with analyte depleted serum for LoQ (tested twelve times over five days).
Analytical specificity (Interference): Three serum samples with different IgG Kappa/Lambda concentrations, spiked with interfering substances.
Antigen Excess Detection: 8 monoclonal IgG Kappa and 6 monoclonal IgG Lambda samples.
Method Comparison (with predicate device):
- IgG Kappa: 284 serum samples (including 135 IgG Kappa paraprotein, 60 IgG Lambda paraprotein, 60 donor samples, and 29 other samples).
- IgG Lambda: 172 serum samples (including 42 IgG Kappa paraprotein, 59 IgG Lambda paraprotein, 59 donor samples, and 1 AL Amyloidosis sample).
- IgG Kappa/Lambda Ratio: 143 serum samples (including 39 IgG Kappa paraprotein, 44 IgG Lambda paraprotein, 59 donor samples, and 1 AL Amyloidosis sample).
Clinical Monitoring Response Category Study: 69 monitoring samples from 10 IgG Kappa patients and 12 IgG Lambda patients. The data provenance is implied to be from patient samples, consistent with clinical monitoring. Country of origin not specified, but typically for submissions to the US FDA, data from well-regulated regions (e.g., US, EU) are accepted. It is a retrospective analysis utilizing existing patient data.
Data Modelling: Monitoring sample results from the original BNII submission (predicate device's clinical data). The total number of evaluations was 437. This is a retrospective application of new data to existing clinical outcomes.
Expected values/Reference range: 50 adult donor samples.

Data Provenance: The document does not explicitly state the country of origin for the clinical samples. The studies are based on patient and donor serum samples. The clinical monitoring study and data modeling are retrospective analyses of previously collected patient data.

3. Number of Experts Used to Establish the Ground Truth for the Test Set and Their Qualifications

This device is an IVD assay, not an AI image analysis device. Therefore, the concept of "experts establishing ground truth for a test set" in the context of image annotation is not directly applicable.

For IVDs, "ground truth" for clinical effectiveness is typically established by:

Reference Methods: Comparison against an established, validated method (the predicate device in this case, the Siemens BN™II).
Clinical Diagnosis/Outcomes: In the context of monitoring multiple myeloma, the "ground truth" for patient response categories (CR, VGPR, PR, SD, PD, Relapse from CR) is defined by clinical guidelines (NCCN Guidelines v1.2011) that integrate multiple clinical tests (immunofixation, bone marrow, urine assessments) and patient history, rather than a single expert adjudication on an image. The study compares the calculated HLC ratio response classification from the new device with that from the predicate based on these clinical definitions.

4. Adjudication Method for the Test Set

Not applicable as this is an IVD assay. The performance is assessed against analytical standards and agreement with a predicate device and established clinical guidelines for patient response classification.

5. If a multi-reader multi-case (MRMC) comparative effectiveness study was done

Not applicable as this is an IVD assay, not an AI-assisted diagnostic tool involving human readers of images.

6. If a standalone (i.e. algorithm only without human-in-the-loop performance) was done

This concept is tailored for AI devices. For this IVD, the "standalone performance" is represented by its analytical performance characteristics (precision, linearity, detection limits, specificity) and its ability to classify patient responses independently using its measured values and interpretation rules, which are then compared to the predicate device's classifications. The device itself (the Optilite analyzer and the kits) provides quantitative measurements.

7. The Type of Ground Truth Used

Analytical Performance: The ground truth for analytical performance (e.g., true concentration for linearity, known interferences for specificity) would typically be established by highly accurate reference methods or certified reference materials.
Clinical Monitoring Response: The ground truth for "clinical response categorization" (e.g., Complete Response, Partial Response) is defined by established clinical guidelines (NCCN Guidelines v1.2011) that consider a combination of laboratory tests (including immunofixation, bone marrow, and urine assessments) rather than a single, independent "ground truth" measure. The study aimed to show agreement between the new device's HLC ratio-based classification and the predicate device's HLC ratio-based classification within these established clinical guidelines.

8. The Sample Size for the Training Set

This document describes a pre-market notification (510(k)) for a medical device that is an in vitro diagnostic quantitative assay. It is not an AI/Machine Learning device that requires a "training set" in the conventional sense for model development. The "training" for such an IVD would be the development and optimization of the assay reagents and analytical procedure by the manufacturer. The data described here are for validation and verification, not for training a machine learning model.

9. How the Ground Truth for the Training Set Was Established

As noted above, there is no "training set" in the context of machine learning for this type of IVD. The development of the assay's performance characteristics (e.g., reagent formulation, calibration curve fitting) would be based on internal development work and validated against established laboratory practices and reference materials.

Summary

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February 15, 2018

Image /page/0/Picture/1 description: The image contains the logo of the U.S. Food and Drug Administration (FDA). The FDA logo consists of two parts: the Department of Health & Human Services logo on the left and the FDA acronym with the full name of the agency on the right. The Department of Health & Human Services logo is a stylized depiction of a human figure, while the FDA part includes the acronym "FDA" in a blue square and the words "U.S. FOOD & DRUG ADMINISTRATION" in blue text.

The Binding Site Group Ltd. Andrea Thomas Regulatory Affairs Specialist 8 Calthorpe Road, Edgbaston Birmingham, B15 1QT Gb

Re: K172613

Trade/Device Name: Optilite Hevylite IgG Kappa Kit, Optilite Hevylite IgG Lambda Kit Regulation Number: 21 CFR 866.5510 Regulation Name: Immunoglobulins A, G, M, D, and E immunological test system Regulatory Class: Class II Product Code: PCN, PCO Dated: August 16, 2017 Received: August 31, 2017

Dear Andrea Thomas:

We have reviewed your Section 510(k) premarket notification of intent to market the device referenced above and have determined the device is substantially equivalent (for the indications for use stated in the enclosure) to legally marketed predicate devices marketed in interstate commerce prior to May 28, 1976, the enactment date of the Medical Device Amendments, or to devices that have been reclassified in accordance with the provisions of the Federal Food, Drug, and Cosmetic Act (Act) that do not require approval of a premarket approval application (PMA). You may, therefore, market the device, subject to the general controls provisions of the Act. The general controls provisions of the Act include requirements for annual registration, listing of devices, good manufacturing practice, labeling, and prohibitions against misbranding and adulteration. Please note: CDRH does not evaluate information related to contract liability warranties. We remind you, however, that device labeling must be truthful and not misleading.

If your device is classified (see above) into either class II (Special Controls) or class III (PMA), it may be subject to additional controls. Existing major regulations affecting your device can be found in the Code of Federal Regulations, Title 21, Parts 800 to 898. In addition, FDA may publish further announcements concerning your device in the Federal Register.

Please be advised that FDA's issuance of a substantial equivalence determination does not mean that FDA has made a determination that your device complies with other requirements of the Act or any Federal statutes and regulations administered by other Federal agencies. You must comply with all the Act's requirements, including, but not limited to: registration and listing (21 CFR Part 807); labeling (21 CFR Part 801 and Part 809); medical device reporting of medical device-related adverse events) (21 CFR 803); good manufacturing practice requirements as set forth in the quality systems (QS) regulation (21 CFR

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Part 820); and if applicable, the electronic product radiation control provisions (Sections 531-542 of the Act); 21 CFR 1000-1050.

Also, please note the regulation entitled, "Misbranding by reference to premarket notification" (21 CFR Part 807.97). For questions regarding the reporting of adverse events under the MDR regulation (21 CFR Part 803), please go to http://www.fda.gov/MedicalDevices/Safety/ReportaProblem/default.htm for the CDRH's Office of Surveillance and Biometrics/Division of Postmarket Surveillance.

For comprehensive regulatory information about medical devices and radiation-emitting products, including information about labeling regulations, please see Device Advice (https://www.fda.gov/MedicalDevices/DeviceRegulationandGuidance/) and CDRH Learn (http://www.fda.gov/Training/CDRHLearn). Additionally, you may contact the Division of Industry and Consumer Education (DICE) to ask a question about a specific regulatory topic. See the DICE website (http://www.fda.gov/DICE) for more information or contact DICE by email (DICE@fda.hhs.gov) or phone (1-800-638-2041 or 301-796-7100).

Sincerely,

Kelly Oliner-S

For, Lea Carrington Director Division of Immunology and Hematology Devices Office of In Vitro Diagnostics and Radiological Health Center for Devices and Radiological Health

Enclosure

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Indications for Use

510(k) Number (if known) K172613

Device Name

Optilite Hevylite IgG Kappa Kit (NK621.OPT) and Optilite Hevylite IgG Lambda Kit (NK622.OPT)

Indications for Use (Describe)

Type of Use (Select one or both, as applicable)
-------------------------------------------------

	X Prescription Use (Part 21 CFR 801 Subpart D)
	Over-The-Counter Use (21 CFR 801 Subpart C)

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Optilite Hevylite Human IgG Kappa and IgG Lambda Kits Submission Summary

A. 510(k) Number:
K172613

B. Purpose for Submission:

Modified IVD assay on previously cleared instrument

C. Measurand:

Immunoglobulin IgG Kappa (combined γ heavy and κ light chain) and Immunoglobulin IgG Lambda (combined y heavy and À light chain)

D. Type of Test:

Quantitative, Turbidimetry

E. Applicant: The Binding Site Group, Ltd.
The Binding Site Group, Ltd.
F. Proprietary and Established Names: Optilite® Hevylite® IgG Kappa Kit Optilite® Hevylite® IgG Lambda Kit

G. Requlatory Information:

1. Regulatory Section: 21 CFR §866.5510, Immunoglobulins A, G, M, D and E Immunological Test System
1. Classification: Class II
1. Product Code:

PCN - IgG Kappa (Heavy and Light chain combined). Antigen, antiserum, control PCO - IgG Lambda (Heavy and Light chain combined). Antigen, antiserum, control

1. Panel:
  Immunology (82)

H. Intended Use:

1. Intended use(s):
  Optilite Hevylite IgG Kappa is a quantitative in vitro assay performed on the Optilite analyser for the measurement of IgG kappa (IgG heavy chain and kappa light chain intact immunoglobulin) in serum. Measurement of Hevylite Human IgG Kappa is used alongside Hevylite Human IgG Lambda to calculate the IgG Kappa / IgG Lambda ratio. The Hevylite Human IgG Kappa / IgG Lambda ratio can be used when monitoring previously diagnosed IgG multiple myeloma and is used in conjunction with other laboratory tests and clinical evaluations. The assignment of complete response is reliant upon other tests including immunofixation, bone marrow and urine assessments.

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Optilite Hevylite IgG Lambda is a quantitative in vitro assay performed on the Optilite analyser for the measurement of IgG lambda (IgG heavy chain and lambda light chain intact immunoglobulin) in serum. Measurement of Hevylite Human IgG Lambda is used alongside Hevylite Human IgG Kappa to calculate the IgG Kappa / IgG Lambda ratio. The Hevylite Human IgG Kappa / IgG Lambda ratio can be used when monitoring previously diagnosed IgG multiple myeloma and is used in conjunction with other laboratory tests and clinical evaluations. The assignment of complete response is reliant upon other tests including immunofixation, bone marrow and urine assessments.

1. Indication(s) for use: Same as intended use.
1. Special conditions for use statement(s): This product is for in vitro diagnostic prescription use only.

Warning: The result of Hevylite Human IgG Kappa in a given specimen determined with assays with different manufacturers and different instrument platforms can vary due to differences in assay methods and reagent specificity. The results reported by the laboratory to the physician must include the identity of the Hevylite Human IgG Kappa assay used. Values obtained with different assay methods cannot be used interchangeably. If, in the course of serially monitoring a patient, the assay method used for determining Hevylite IgG Kappa levels is changed, additional sequential testing should be carried out. Prior to changing assays, the laboratory MUST confirm baseline values for patients being serially monitored.

Warning: The result of Hevylite Human IgG Lambda in a given specimen determined with assays with different manufacturers and different instrument platforms can vary due to differences in assay methods and reagent specificity. The results reported by the laboratory to the physician must include the identity of the Hevylite Human IgG Lambda assay used. Values obtained with different assay methods cannot be used interchangeably. If, in the course of serially monitoring a patient, the assay method used for determining Hevylite IgG Lambda levels is changed, additional sequential testing should be carried out. Prior to changing assays, the laboratory MUST confirm baseline values for patients being serially monitored.

1. Special instrument requirements: The Binding Site Optilite analyser

Device Description: l.

J. Substantial Equivalence Information:

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1. Predicate device names and predicate 510(k) number: Hevylite Human IgG Kappa Kit and Hevylite Human IgG Lambda Kit for use on the Siemens BN™II (K132555)
1. Comparison with predicate:

Similarities
Item	Device	Predicate
Intended Use	Quantitative in vitro assay for themeasurement of IgG Kappa (IgGheavy chain and kappa light chainintact immunoglobulin) and IgGLambda (IgG heavy chain andlambda light chain intactimmunoglobulin) in serum.Measurement of Hevylite Human IgGKappa is used alongside HevyliteHuman IgG Lambda to calculate theIgG Kappa/IgG Lambda ratio. TheHevylite Human IgG Kappa/IgGLambda ratio can be used whenmonitoring previously diagnosed IgGmultiple myeloma and is used inconjunction with other laboratorytests and clinical evaluations. Theassignment of complete response isreliant upon other tests includingimmunofixation, bone marrow andurine assessments.	Same
Analyte	IgG Kappa and Lambda	Same
Antibody	Polyclonal monospecific sheep anti-human combined γ heavy and κ lightchain or combined γ heavy and λlight chain.	Same
Control	Binding Site High and Low Controls	Same
Traceability	DA470k	Same
SampleMatrix	Serum	Same
CaptureAntibody	Sheep anti-human IgG combined	Same
Calibrator	Single level Binding Site Hevylitecalibrator autodiluted by the analyserto six different concentrations	Same
Open VialStability	3 months	Same
ReferenceInterval	IgGK: 4.03 - 9.78 g/LIgGL: 1.97 - 5.71 g/LIgGK/IgGL Ratio: 0.98 - 2.75	Same

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Differences
Item	Device	Predicate
Method	Turbidimetry	Nephelometry
Instrument	Binding Site Optilite	Siemens BN™ II Systems
On BoardStability	28 days	Not stated
MeasuringRange	IgGK: 2.3 – 30.0 g/L (1+19 dilution)IgGL: 1.5 – 17.5 g/L (1+19 dilution)	IgGK: 1.72 – 27.5 g/L (1/100 dilution)IgGL: 0.88 – 14.0 g/L (1/100 dilution)
	Extended Range for IgGK:1+0 dilution: 0.115 – 1.5 g/L1+4 dilution: 0.575 – 7.5 g/L1+79 dilution: 9.2 - 120 g/L	Extended Range for IgGK:1/5 dilution: 0.086 – 1.375 g/L1/20 dilution: 0.344 – 5.500 g/L1/400 dilution: 6.88 – 110.0 g/L1/2000 dilution: 34.4 – 550 g/L
	Extended Range for IgGL:1+0 dilution: 0.075 – 0.875 g/L1+4 dilution: 0.375 – 4.375 g/L1+79 dilution: 6 - 70 g/L1+119 dilution: 9 – 105 g/L	Extended Range for IgGL:1/5 dilution: 0.044 – 0.700 g/L1/20 dilution: 0.175 – 2.800 g/L1/100 dilution: 0.88 – 14.00 g/L1/400 dilution: 3.50 – 56.0 g/L1/2000 dilution: 17.5 – 280 g/L

K. Standard/Guidance Documents Referenced (if applicable):

CLSI EP17-A Protocols for Determination of Limits of Detection and Limits of Quantitation; Approved Guideline.

CLSI EP7-A2 Interference Testing in Clinical Chemistry, Approved Guideline -Second Edition.

CLSI EP6-A: Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach.

CLSI EP5-A2 Evaluation of Precision Performance of Quantitative Measurement Methods; Approved Guideline - Second Edition.

CLSI C28-A3: Defining, Establishing and Verifying Reference Intervals in the Clinical Laboratory.

L. Test Principle:

Evaluating the concentration of a soluble antigen (e.g. IgG lambda) by turbidimetry involves the addition of the test sample to a solution containing the appropriate antibody (anti-IgG lambda) in a reaction vessel or cuvette. A beam of light is passed through the cuvette and, as the antigen-antibody reaction proceeds, the light passing through the cuvette is scattered increasingly as insoluble immune complexes are formed. Light scatter is monitored by measuring the decrease in intensity of the incident beam of light. The antibody in the cuvette is in excess so the amount of immune complex formed is proportional to the antigen concentration. A series of calibrators of known antigen concentration are assayed initially to produce a calibration curve of measured light scatter versus antigen concentration. Samples of unknown antigen concentration can then be assayed and the results read from the calibration curve.

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M. Performance Characteristics (if/when applicable):

1. Analytical performance:

a. Precision/Reproducibility:
The within-run, between-run, between-day, between-lot and betweeninstrument precision were determined by testing six serum samples over 21 days with two runs per day on three different reagent lots on three analysers. Results are summarised below.

Sample	Mean	Within Run		BetweenRun		BetweenDay		BetweenLot		BetweenInstrument		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1	4.260	0.184	4.3	0.209	4.9	0.162	3.8	0.140	3.3	0.033	0.8	0.323	7.6
2	3.148*	0.048	1.5	0.104	3.3	0.124	3.9	0.098	3.1	0.036	1.1	0.169	5.4
3	5.601*	0.145	2.6	0.263	4.7	0.308	5.5	0.205	3.7	0.099	1.8	0.430	7.7
4	7.621	0.178	2.3	0.263	3.5	0.413	5.4	0.426	5.6	0.216	2.8	0.521	6.8
5	13.574	0.218	1.6	0.385	2.8	0.879	6.5	0.851	6.3	0.253	1.9	0.984	7.2
6	16.824	0.383	2.3	0.352	2.1	0.992	5.9	0.991	5.9	0.608	3.6	1.120	6.7

IqGK Precision studies:

Sample	Mean	Within Run		BetweenRun		BetweenDay		BetweenLot		BetweenInstrument		Total
		SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV	SD	%CV
1	2.894	0.118	4.1	0.120	4.2	0.198	6.8	0.119	4.1	0.142	4.9	0.260	9.0
2	1.313*	0.029	2.2	0.034	2.6	0.068	5.2	0.054	4.1	0.037	2.8	0.081	6.2
3	2.379*	0.059	2.5	0.049	2.1	0.064	2.7	0.061	2.6	0.027	1.1	0.100	4.2
4	4.262	0.096	2.2	0.144	3.4	0.267	6.3	0.182	4.3	0.190	4.4	0.318	7.5
5	6.657	0.138	2.1	0.112	1.7	0.301	4.5	0.256	3.8	0.200	3.0	0.350	5.3
6	13.941	0.297	2.1	0.237	1.7	0.414	3.0	0.541	1.7	0.096	0.7	0.561	4.0

sample run at 1+4 dilution.

b. Linearity/assay reportable range:
A linearity study was performed following CLSI document Evaluation of the Linearity of Quantitative Measurement Procedures: A Statistical Approach. The linearity of the IgGK and IgGL assays have been confirmed using serially diluted serum samples to cover the standard measuring ranges of 2.3 - 30.0 q/L and 1.5 - 17.5 g/L respectively. The results demonstrated that the IqGK and IgGL assays are linear over the ranges of 1.930 - 33.427 g/L and 1.380 -19.300 g/L at 1+19 dilution with deviation from linearity ≤ 10%.
c. Traceability, Stability, Expected values (controls, calibrators, or methods): Traceability: An Internal Reference material (IR) was assigned by comparison with the Reference Material DA470K.

Stability:

Open-vial stability was performed on three lots of Optilite Hevylite IgG Kappa and IgG Lambda kits with testing intervals at various time points up to 112 days. Data supports an open vial stability claim of 3 months at 2-8°C.

On-board stability was performed on three lots of Optilite Hevylite IgG Kappa and IgG Lambda kits with testing intervals at various time points up to 37 days for IgGK and

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49 days for IgGL. Data supports an on-board stability claim of 28 days, provided that the power is left switched on as stated in the product insert.

d. Detection limit:
The analytical sensitivity was determined in accordance with CLSI EP17-A. The Limit of Blank was based on 60 determinations of analyte depleted sample and was estimated at the 95th percentile of the distribution. The Limit of Detection was calculated from the LoB and the combined SD of the 5 LoQ samples. The LoQ was calculated from 5 independent samples (serum samples diluted with analyte depleted serum to achieve a concentration close to the bottom of the measuring range) tested twelve times over five days. The tabulated summary of results is shown below:

	LoB	LoD	LoQ
IgG Kappa	0.000 g/L	0.009 g/L	0.115 g/L
IgG Lambda	0.000 g/L	0.005 g/L	0.075 g/L

e. Analytical specificity:

Interference:

Interferences were assessed according to CLSI EP7-A2 by testing three serum samples with different IgG Kappa and IgG Lambda concentration ranges, including a sample close to the lower limit of the reference interval, a sample within the reference interval and an elevated serum sample. Each sample was spiked with interfering substances and tested in multiple replicates. The acceptance criterion was that the mean results from the spiked samples must be within ±10% of the mean of the control samples.

No significant assay interference effects were observed when the samples were tested with bilirubin at 200mg/L, haemoglobin at 5g/L, triglyceride at 1000mg/dL. intralipid at 125mg/dL or the 16 commonly used drugs at the concentrations given below.

Substance	Concentration
Acetaminophen	1324µmol/L
Acetylsalicylic Acid	3.63mmol/L
Ascorbic Acid	342µmol/L
Bortezomib	6mg/mL
Caffeine	308µmol/L
Cimetidine	79.2µmol/L
Cyclophosphamide Monohydrate	60µg/mL
Digoxin	3.9nmol/L (IgGK)7.8nmol/L (IgGL)
Furosemide	181µmol/L
Ibuprofen	2425µmol/L
Methotrexate	2mmol/L
Penicillin	75mg/L
Phenytoin	198µmol/L
Pomalidomide	100µg/mL
Prednisolone	100µg/mL
Theophylline	222µmol/L

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Cross reactivity:

No significant cross reaction was observed during testing for the predicate device. The specificity of the antisera is unchanged.

Antigen Excess Detection:

The possibility of antigen excess occurring when using the device on the Binding Site Optilite was evaluated with 8 monoclonal IgG Kappa and 6 monoclonal IgG Lambda samples with concentrations above the respective standard measuring ranges. No antigen excess was observed up to 100.5q/L and 102.5g/L for IgG Kappa and IgG Lambda respectively.

f. Assay cut-off:
Refer to Expected values.

2. Comparison studies:

a. Method comparison with predicate device:

lqG Kappa:

A comparison study was performed by analysing 284 serum samples (including 135 IgG Kappa paraprotein and 60 IgG Lambda paraprotein samples, 60 donor samples and 29 other samples, covering the range 0.2 -47.69 g/L) using the Optilite Hevylite IgG Kappa kit and an alternative commercially available assay. Passing Bablok regression analysis generated the following results:

y = 1.10x - 0.68 g/L correlation coefficient r = 0.957 (y = Optilite, x = predicate analyser)

lgG Lambda:

A comparison study was performed by analysing 172 serum samples (including 42 IqG Kappa paraprotein and 59 IgG Lambda paraprotein samples, 59 donor samples and 1 AL Amyloidosis sample, covering the range 0.09 - 40.65 q/L) using the Optilite Hevvlite IgG Lambda kit and an alternative commercially available assay. Passing Bablok regression analysis generated the following results:

(y = Optilite, x = predicate analyser) y = 0.95x + 0.00 g/L correlation coefficient r = 0.823

lgG Kappa/Lambda Ratio:

A comparison study was performed by analysing 143 serum samples (including 39 IgG Kappa paraprotein and 44 IgG Lambda paraprotein samples, 59 donor samples and 1 AL Amyloidosis sample, covering the range 0.01 - 277.50 g/L) using the Optilite Hevylite IgG Kappa and IgG Lambda kits and alternative commercially available assays. Passing Bablok regression analysis generated the following results:

(v = Optilite, x = predicate analyser) v = 1.12x - 0.17 correlation coefficient r = 0.901

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b. Matrix comparison: Not applicable
1. Clinical studies:
- a. Clinical Sensitivity/clinical specificity: Transformation of the BNII Study onto the Optilite

Purpose of the study

The purpose of this study was to compare the clinical HLC Response categorisation of the predicate and the Optilite Hevylite IgG Kappa and Lambda Kits obtained from samples taken at multiple time points from IgG Kappa and IgG Lambda multiple myeloma patients during the course of their disease. The sponsor generated Passing and Bablok regression equations for the comparison study of the Optilite kits against the predicate kits. The regression equation was then modelled with the existing monitoring sample results from the original BNII submission to evaluate the clinical validity of the new device.

Complete Response(CR)	HLC ratio within the normal range and negative urine immunofixation.
Very Good PartialResponse (VGPR)	>91% reduction of HLC ratio from baseline and reduction in 24 hoururinary M-protein to ≤100mg per 24 hours
Partial Response(PR)	Reduction of HLC ratio from baseline between 47 - 91% and reduction in24 hours urinary M-protein by ≥90% or to ≤200mg/24 hours.
Stable Disease (SD)	A change in HLC ratio from baseline < 32% increase but < 47% reduction.
Progressive Disease(PD)	> 32% increase in HLC ratio from baseline (the absolute increase ininvolved IgG must be ≥5g/L) or a ≥25% increase in urine M-protein frombaseline (the absolute increase must be ≥200mg/24 hours)
Relapse from CR	> 32% increase in HLC ratio from baseline (the absolute increase ininvolved IgG must be ≥5g/L)

Table 1: HLC Monitoring Response Category

Optilite HLC Response Category Study

Assignment of classification was based on the HLC Monitoring Response Category detailed in Table 1, using all assay data available. Responses were cateqorised in accordance with NCCN Guidelines v1.2011 by using the percentage change from baseline. Responses were characterised as progressive disease (PD), stable disease (SD), partial response (PR), very good partial response (VGPR), and complete response (CR). Kappa statistics were used to evaluate agreement between the test and predicate devices.

Optilite Monitoring Study Design

A comparison of 69 monitoring samples from 10 IqG Kappa patients and 12 IgG Lambda patients was performed to compare the BNII HLC response category assigned to those observed with the Optilite Hevylite IgG Kappa and Lambda kits (Note: The Optilite monitoring response category is not to be used interchangeably with other manufacturer's assays or with any other instrument platform monitoring response category). The results of the comparison study using 69 monitoring samples vielding 43 response classifications are shown in the table below:

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Observed		Predicate HLC response
		CR	VGPR	PR	SD	PD	Total
Optilite HLC Response	CR	0	0	0	0	0	0
	VGPR	0	1	0	0	0	1
	PR	0	0	11	1	0	12
	SD	0	0	1	23	1	25
	PD	0	0	0	2	3	5
Total		0	1	12	26	4	43
Kappa (95% CIs)		0.79 (0.62 – 0.96)
Weighted Kappa (95% CIs)		0.87 (0.74 – 0.98)

These monitoring data were also supported by additional statistical regression equation modelling data.

Data Modelling:

A data modelling procedure was carried out on monitoring sample results from the original BNII submission. The Passing Bablok regression equations derived from the comparison study and the total imprecision values derived from the precision studies were used to mathematically transform the data. The results are summarised in the tables below.

H/L*		Predicate Assigned Response
		CR	VGPR	PR	SD	PD	Total
TransformedData Response	CR	29	5	2	0	0	36
	VGPR	24	72	27	0	0	123
	PR	7	4	165	4	0	180
	SD	0	1	4	92	0	97
	PD	0	0	0	0	1	1
Total		60	82	198	96	1	437
Kappa (95% CIs)		0.75 (0.70 – 0.80)
Weighted Kappa (95% CIs)		0.86 (0.82 – 0.89)

H/L: H = highest kappa, L = lowest lambda; the imprecision values were applied to generate the highest possible kappa and the lowest possible lambda results.

L/H**		Predicate Assigned Response
		CR	VGPR	PR	SD	PD	Total
TransformedData Response	CR	55	16	6	1	0	78
	VGPR	5	63	23	0	0	91
	PR	0	2	166	3	0	171
	SD	0	1	3	92	0	96
	PD	0	0	0	0	1	1
Total		60	82	198	96	1	437
Kappa (95% CIs)		0.81 (0.76 - 0.85)
Weighted Kappa (95% CIs)		0.90 (0.86 - 0.93)

** L/H: L = lowest kappa, H = highest lambda; the imprecision values were applied to generate the lowest possible kappa and the highest possible lambda results.

b. Other clinical supportive data (when a. is not applicable) Not applicable.

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4. Clinical cut-off:

Refer to discussion above.

1. Expected values/Reference range:
  The reference intervals were transferred from the predicate devices and were verified by testing 50 adult donor samples.

	95 Percentile Range
IgG kappa (g/L)	4.03 – 9.78 g/L
IgG lambda (g/L)	1.97 - 5.71 g/L
IgG kappa/ IgG lambda ratio	0.98 - 2.75

N. Proposed Labelling:

The labelling is sufficient and it satisfies the requirements of 21 CFR Part 809.10.

O. Conclusion:

The submitted information in this premarket notification is complete and supports a substantial equivalence decision.

Regulation Number and Section

§ 866.5510 Immunoglobulins A, G, M, D, and E immunological test system.

(a)
Identification. An immunoglobulins A, G, M, D, and E immunological test system is a device that consists of the reagents used to measure by immunochemical techniques the immunoglobulins A, G, M, D, an E (serum antibodies) in serum. Measurement of these immunoglobulins aids in the diagnosis of abnormal protein metabolism and the body's lack of ability to resist infectious agents.(b)
Classification. Class II (performance standards).